Cancer preventive bioactive food compounds are well documented. A clear understanding of the molecular targets for these food components is fundamental to the development of effective diet strategies against cancer. Despite the development of many methods, target identification for bioactive compounds is an unmet challenge due to the huge diversity in small-molecule structure, activity, and mechanisms of action. Current affinity-based target identification techniques are limited by the necessity to modify each drug individually, while indirect, non-affinity based approaches are dependent on the drug's ability to induce specific biochemical or cellular readouts. This project presents a novel, universally applicable target identification approach (DARTS) that analyzes direct small-molecule binding to its protein target without requiring modification or immobilization of the small molecule. The specific aims are to 1) establish a robust DARTS protocol for identifying the molecular targets of bioactive food components, and 2) to use DARTS to determine the molecular targets of bioactive food compounds that explain their inhibition of human cancer cell growth. If successful, the studies will have a major impact on our understanding of the linkage between diet and cancer, and shed light on new mechanisms and targets for cancer prevention and aging intervention. PUBLIC HEALTH RELEVANCE: Cancer preventive agents in our food are well documented. A clear understanding of the molecular targets for these bioactive food components is challenging but fundamental to the development of effective diet strategies against cancer. This project develops a novel and universal method to identify the molecular targets for anti-cancer food components.